Human PLUNC Antibody

Detection of PLUNC by Western Blot

JNK/c-Jun pathway is involved in LPS-mediated up-regulation of BPIFA1 expression in nasal epithelial cells.(A) Cells were pretreated for 1 h with 20 μM PD98059 (ERK inhibitor), 10 μM SP600125 (JNK inhibitor), or 20 μM SB203580 (p38 inhibitor) and incubated with 10 μg/ml LPS for 2 h. (B) Cells were transfected with a JNK-dominant negative (DN-JNK) mutant for 24 h or pretreated with SP600125 for 30 min prior to incubation with LPS for 1 h. The protein expression levels were determined using western blot. beta-actin was used as the loading control. The western blots were carried out independently in triplicate and results were representative of one of three independent experiments. The expression level of each protein was quantified by signal intensity and was indicated at the bottom of each lane. The quantitative analysis of western blot for three independent experiments was shown in S2 Fig. ANOVA with Tukey’s test was used to compare the overall difference between the groups. P < 0.05 was considered statistically significant. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/26646664), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of PLUNC by Western Blot

AP-1 is involved in LPS-induced BPIFA1 expression. Cells were pretreated with 10 μM curcumin or tanshinone (inhibitors of c-Jun) for 30 min, followed by incubation with LPS (10 μg/ml) for 2 h. Protein expression levels were determined using western blot and normalized to those of beta-actin. The expression level of each protein is quantified by signal intensity and is indicated at the bottom of each lane. The quantitative analysis of western blot for three independent experiments was shown in S3 Fig. ANOVA with Tukey’s test was used to compare the overall difference between the groups. P < 0.05 was considered statistically significant. (B) Cells were transfected with AP-1-Luc reporter and incubated with LPS (10 μg/ml) for another 2 h. Cell lysates were subjected to luciferase activity assays to determine AP-1 luciferase activity. The results represented mean and standard deviation values from three independent experiments. *, P < 0.05 compared between two groups. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/26646664), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of PLUNC by Western Blot

JNK/c-Jun pathway is involved in LPS-mediated up-regulation of BPIFA1 expression in nasal epithelial cells.(A) Cells were pretreated for 1 h with 20 μM PD98059 (ERK inhibitor), 10 μM SP600125 (JNK inhibitor), or 20 μM SB203580 (p38 inhibitor) and incubated with 10 μg/ml LPS for 2 h. (B) Cells were transfected with a JNK-dominant negative (DN-JNK) mutant for 24 h or pretreated with SP600125 for 30 min prior to incubation with LPS for 1 h. The protein expression levels were determined using western blot. beta-actin was used as the loading control. The western blots were carried out independently in triplicate and results were representative of one of three independent experiments. The expression level of each protein was quantified by signal intensity and was indicated at the bottom of each lane. The quantitative analysis of western blot for three independent experiments was shown in S2 Fig. ANOVA with Tukey’s test was used to compare the overall difference between the groups. P < 0.05 was considered statistically significant. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/26646664), licensed under a CC-BY license. Not internally tested by R&D Systems.